	US 12,134,633 B2
	Separation matrix
Gustav José Rodrigo, Uppsala (SE); Tomas Bjorkman, Uppsala (SE); Mats Ander, Uppsala (SE); and Jesper Ulf Hansson, Uppsala (SE)
Assigned to Cytiva BioProcess R&D AB, Uppsala (SE)
Filed by Cytiva BioProcess R&D AB, Uppsala (SE)
Filed on Apr. 11, 2023, as Appl. No. 18/298,857.
Application 18/298,857 is a continuation of application No. 17/221,438, filed on Apr. 2, 2021, granted, now 11,685,764.
Application 17/221,438 is a continuation of application No. 16/893,574, filed on Jun. 5, 2020, granted, now 10,995,113, issued on May 4, 2021.
Application 16/893,574 is a continuation of application No. 16/682,855, filed on Nov. 13, 2019, granted, now 10,711,035, issued on Jul. 14, 2020.
Application 16/682,855 is a continuation of application No. 16/096,952, granted, now 10,513,537, issued on Dec. 24, 2019, previously published as PCT/EP2017/061164, filed on May 10, 2017.
Application 16/096,952 is a continuation of application No. 15/282,367, filed on Sep. 30, 2016, granted, now 10,654,887, issued on May 19, 2020.
Claims priority of application No. 1608229 (GB), filed on May 11, 2016; and application No. 1608232 (GB), filed on May 11, 2016.
Prior Publication US 2023/0279047 A1, Sep. 7, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 1/22 (2006.01); B01D 15/38 (2006.01); B01J 20/24 (2006.01); B01J 20/28 (2006.01); B01J 20/286 (2006.01); B01J 20/32 (2006.01); C07K 14/31 (2006.01); C07K 16/00 (2006.01); C07K 16/06 (2006.01); C07K 17/10 (2006.01)

CPC C07K 1/22 (2013.01) [B01D 15/3809 (2013.01); B01J 20/24 (2013.01); B01J 20/28016 (2013.01); B01J 20/28078 (2013.01); B01J 20/286 (2013.01); B01J 20/3212 (2013.01); B01J 20/3274 (2013.01); C07K 14/31 (2013.01); C07K 16/00 (2013.01); C07K 16/065 (2013.01); C07K 17/10 (2013.01)]

38 Claims

1. A separation matrix comprising at least 15 mg/ml Fc-binding ligands covalently coupled to a porous support, wherein:

the ligands comprise multimers of alkali-stabilized Protein A domains;

the porous support comprises cross-linked polymer particles having a volume-weighted median diameter (d50,v) of from 56 μm to 70 μm;

the separation matrix has a max pressure of at least 0.58 MPa when packed at 300+/−10 mm bed height in a 35 mm separation column; and

the Protein A domains comprise an Fc-binding polypeptide defined by, or having at least 80% identity to SEQ ID NO: 53 and

X₁QX₂AFYEILX₃LPNLTEEQRX₄X₅FIX₆X₇LKDX₈PSX₉SX₁₀X₁₁X₁₂LAEAKX₁₃X₁₄N X₁₅AQ (SEQ ID NO: 53), wherein individually of each other:

X₁=A or Q or is deleted;

X₂=E, K, Y, T, F, L, W, I, M, V, A, H, or R;

X₃=H or K;

X₄=A or N;

X₅=A, G, S, Y, Q, T, N, F, L, W, I, M, V, D, E, H, R, or K;

X₆=Q or E;

X₇=S or K;

X₈=E or D;

X₉=W or V or is deleted;

X₁₀=K, R, or A, or is deleted;

X₁₁=A, E, or N, or is deleted;

X₁₂=I or L;

X₁₃=K or R;

X₁₄=L or Y; and

X₁₅=D, F, Y, W, K, or R.